Phylogenomics, taxonomy and morphological characters of the Microdochiaceae (Xylariales, Sordariomycetes)

683 MycoKeys

MycoKeys 106: 303-325 (2024)
DOI: 10.3897/mycokeys.106.127355

Research Article

Phylogenomics, taxonomy and morphological characters of the
Microdochiaceae (Xylariales, Sordariomycetes)

Zhao-Xue Zhang™, Yu-Xin Shang', Meng-Yuan Zhang|, Jin-Jia Zhang', Yun Geng?,

Ji-Wen Xia'™®, Xiu-Guo Zhang’

1 Shandong Provincial Key Laboratory for Biology of Vegetable Diseases and Insect Pests, College of Plant Protection, Shandong Agricultural University, Taian,

271018, China

2 Institute of Crop Germplasm Resources, Shandong Academy of Agricultural Sciences, Jinan, 250100, China
Corresponding author: Xiu-Guo Zhang (sdau613@163.com)

This article is part of:

Exploring the Hidden Fungal Diversity:
Biodiversity, Taxonomy, and Phylogeny
of Saprobic Fungi

Edited by Samantha C. Karunarathna,
Danushka Sandaruwan Tennakoon,
Ajay Kumar Gautam

OPEN Qaceess

Academic editor:

Danushka Sandaruwan Tennakoon
Received: 12 May 2024

Accepted: 20 June 2024

Published: 3 July 2024

Citation: Zhang Z-X, Shang Y-X, Zhang
M-Y, Zhang J-J, Geng Y, Xia J-W,
Zhang X-G (2024) Phylogenomics,
taxonomy and morphological
characters of the Microdochiaceae
(Xylariales, Sordariomycetes).
Mycokeys 106: 303-325. https://doi.
org/10.3897/mycokeys.106.127355

Copyright: © Zhao-Xue Zhang et al.
This is an open access article distributed under
terms of the Creative Commons Attribution

License (Attribution 4.0 International - CC BY 4.0).

Abstract

Species of the family Microdochiaceae (Xylariales, Sordariomycetes) have been reported
from worldwide, and collected from different plant hosts. The proposed new genus and
two new species, viz., Macroidriella gen. nov., M. bambusae sp. nov. and Microdochium
australe sp. nov., are based on multi-locus phylogenies from a combined dataset of ITS
rDNA, LSU, RPB2 and TUB2 with morphological characteristics. Microdochium sinense
has been collected from diseased leaves of Phragmites australis and this is the first
report of the fungus on this host plant. Simultaneously, we annotated 10,372 to 11,863
genes, identified 4,909 single-copy orthologous genes, and conducted phylogenomic
analysis based on genomic data. A gene family analysis was performed and it will expand
the understanding of the evolutionary history and biodiversity of the Microdochiaceae.
The detailed descriptions and illustrations of species are provided.

Key words: Microdochiaceae, multigene phylogeny, new taxa, phylogenomics, taxonomy

Introduction

Microdochium Syd. & P. Syd., is the type genus of the family Microdochiaceae
Hern.-Restr., Crous & J.Z. Groenew. This was first described by Syd. & P. Syd.
(Sydow 1924). The holotype collection of the type species of Microdochium,
M. phragmitis Syd. & P. Syd. was obtained in Germany from the leaves of
Phragmites australis (Sydow 1924). Microdochium species were collected as
endophytes, plant pathogens, and saprophytes, and were frequently isolated
from different plant hosts (Von Arx 1987; Glynn et al. 2005; Jewell and Hsiang
2013; Mandyam et al. 2013; Hiruma et al. 2018; Liang et al. 2019; Lu et al. 2023;
Zhang et al. 2023a). Prior research has indicated that the classification of
Microdochium within the Amphisphaeriaceae is supported by its morphological
similarities (Parkinson et al. 1981; Samuels and Hallett 1983; Von Arx 1984;
Jaklitsch and Voglmayr 2012). Hernandez-Restrepo et al. (2016) was proposed
that /driella and Microdochium may be closely related genera. Their phylogenetic
analysis revealed that /driella, Microdochium, and Selenodriella formed a distinct

303

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

monophyletic group within the Xylariales. Therefore, Hernandez-Restrepo et al.
(2016) established the new family Microdochiaceae to encompass this clade.

Currently, there are approximately 68 species of Microdochium listed in the Index
Fungorum (2024), with 45 species being accepted. Microdochium has a diverse
range of hosts that are widely distributed worldwide (Zhang et al. 2017; Crous et al.
2018, 2019, 2021; Marin-Felix et al. 2019; Huang et al. 2020). However, only a few
species of Microdochium have the capability to cause diseases, primarily impact-
ing grasses and cereals. Zhang et al. (2015) identified Microdochium paspali Syd.
& P. Syd., which was responsible for causing leaf blight on Paspalum vaginatum
Sw. Liang et al. (2019) identified Mi. poae J.M. Liang & Lei Cai, which induced leaf
blight disease in turf grasses like Poa pratensis and Agrostis stolonifera L. Stewart
et al. (2019) identified Mi. sorghi U. Braun, which was responsible for the develop-
ment of zonate leaf spots and decay on sorghum species. Mi. albescens (Thiim.)
Hern.-Restr. & Crous was the causative agent of leaf scald and grain discoloration
in rice, leading to a global decrease in rice yield (Dirchwolf et al. 2023). Mi. bolleyi
(R. Sprague) de Hoog & Herm.-Nijh. was cited as the cause of root necrosis and
basal rot in creeping bent grass (Hong et al. 2008). In addition to this, some species
of Microdochium occur as endophytes or saprophytes. Liu et al. (2022) identified
three species, Microdochium miscanthi S.B. Liu, X.Y. Liu, Z. Meng & X.G. Zhang,
Mi. sinense S.B. Liu, X.Y. Liu, Z. Meng & X.G. Zhang, and Mi. hainanense S.B. Liu, X.Y.
Liu, Z. Meng & X.G. Zhang, isolated from Miscanthus sinensis Anderss. and Phrag-
mites australis (Cav.) Trin. ex Steud in Hainan, China. Zhang et al. (2023a) collected
novel species (Mi. bambusae J. Zhang, Z.X. Zhang, & Z. Li, Mi. nannuoshanense J.
Zhang, Z.X. Zhang, & Z. Li, and Mi. phyllosaprophyticum J. Zhang, Z.X. Zhang, & Z.
Li) from leaves of Bambusaceae plant as a saprobe.

With the advent of the sequencing era, genomics is increasingly being utilized
for phylogenetic studies and can offer additional insights into pathogenic mecha-
nisms (Manamgoda et al. 2011; Schoch et al. 2012; Jeewon et al. 2013; David et al.
2016; Mesny et al. 2021; Tsers et al. 2023). However, at present, only the genome
information of three species of this taxon (Microdochium) can be retrieved from
the NCBI database (https://www.ncbi.nlm.nih.gov/, accessed on 30 April 2024).
In this study, we explored the species diversity of Microdochium and described
one new species and one new host record based on the molecular phylogenet-
ic analyses and morphological observations. In addition, we conducted genome
and transcriptome sequencing of the new species, aiming to conduct phyloge-
netic analysis, and gene structure annotation at the genomic level. By comparing
and analyzing the obtained data with existing species genome information, we
aim to reveal the genetic relationship and functional differences between the new
species and other species. This will gain a more comprehensive understanding of
the biological characteristics and evolutionary history of the new taxa.

Materials and methods
Morphological study

During a series of field visits in 2023 in Hainan Province, China, plant spec-
imens with necrotic spots were collected. Even though specimens harbor
multiple fungi, we managed to obtain pure colonies through the single spore

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 304

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

isolation (Senanayake et al. 2020) and tissue isolation techniques (Zhang et
al. 2023a). We retrieved small fragments (5 x 5 mm) from the damaged leaf
edges, treated them by immersion in a 75% ethanol solution for 60 s, followed
by rinsing in sterile distilled water for 45 s and a 10% sodium hypochlorite
solution for 45 s. Subsequently, specimens were rinsed three times in ster-
ile deionized water for 30 s. The processed fragments were then placed on
Sterile filter paper to remove excess moisture before being transferred onto
PDA for incubation at 24 °C for 3 days. The hyphal tips from growing colonies
were transferred to fresh PDA plates. Images were captured using a Sony Al-
pha 6400L digital camera (Sony Group Corporation, Tokyo, Japan) on days 7
and 14. Microscopic examination of the fungal structures was conducted us-
ing an Olympus SZ61 stereo microscope and an Olympus BX43 microscope
(Olympus Corporation, Tokyo, Japan), along with BioHD-A20c color digital
camera (FluoCa Scientific, China, Shanghai) for recording. All fungal strains
were preserved in 15% sterilized glycerol at 4 °C, with each strain stored in
three 2.0 mL tubes for future studies. Structural measurements were carried
out using Digimizer software (v5.6.0), with a minimum of 25 measurements
taken for each characteristic such as conidiophores, conidiogenous cells, and
conidia. Specimens were deposited in the HSAUP (Herbarium of Plant Pathol-
ogy, Shandong Agricultural University) and HMAS (Herbarium Mycologicum
Academiae Sinicae), while living cultures were stored in the SAUCC (Shan-
dong Agricultural University Culture Collection) for preservation and further
research purposes. Taxonomic information of the new taxa was submitted to
MycoBank (http://www.mycobank.org).

DNA extraction, amplification and sequencing

Fungal DNA was extracted from fresh mycelia grown on PDA using either
the CTAB method or a kit method (OGPLF-400, GeneOnBio Corporation,
Changchun, China) (Guo et al. 2000; Zhang et al. 2023a). Four gene regions,
LSU, ITS, RPB2, and TUB2 were amplified using the primer pairs listed in
Suppl. material 1 (Vilgalys et al. 1990; White et al. 1990; Liu et al. 1999;
Sung et al. 2007; Jewell et al. 2013). The amplification reaction was con-
ducted in a 25 uL reaction volume, consisting of 12.5 uL 2 x Hieff Canace®
Plus PCR Master Mix (Shanghai, China) (with dye) (Yeasen Biotechnology,
Cat No. 10154ES03), 0.5 uL each of forward and reverse primer, and 0.5 uL
template genomic DNA, with the volume adjusted to 25 uL using distilled de-
ionized water. PCR products were separated and purified using 1% agarose
gel and GelRed (TsingKe, Qingdao, China), and UV light was used to visual-
ize the fragments. Gel extraction was performed using a Gel Extraction Kit
(Cat: AE0101-C) (Shandong Sparkjade Biotechnology Co., Ltd., Jinan, Chi-
na). The purified PCR products were subjected to bidirectional sequencing
by Biosune Company Limited (Shanghai, China). The raw data (trace data)
were analyzed using MEGA v. 7.0 to obtain consistent sequences (Kumar et
al. 2016). All sequences generated in this study were deposited in GenBank
under the accession numbers provided in Table 1. The abbreviations of the
genera names used in our study are as follows: /. = Idriela; S. = Selenodriella;
Ma. = Macroidriella; Mi. = Microdochium.

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 305

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Table 1. GenBank accession number of the taxa used in phylogenetic reconstruction.

Species

Cryptostroma corticale

Idriela lunata

|. chlamydospora

|. multiformispora

Macroidriella
bambusae

Microdochium
albescens
Mi. australe

Mi. bambusae

Mi. bolleyi

=

i. chrysanthemoides

=

i. chrysopogonis

=

ij. chuxiongense

=

i, citrinidiscum

=

i, colombiense

=

i, dawsoniorum

=

i. fisheri

S

i, graminearum

S

i, hainanense

S

i, indocalami

S

i, insulare

S

ij. lycopodinum

S

i, maculosum

S

i, Majus

S

i, miscanthi

=

i; Musae

=

i; nannuoshanense

Strain no.

CBS 218.52
CBS 204.56*
CBS 177.57

CGMCC 3.20778*

GZUIFR 21.922
CGMCC 3.20779*
GZUIFR 21.924
GZUIFR 21.925
SAUCC 6792-1*
SAUCC 6792-2
SAUCC 6792-5
SAUCC 6113-1
SAUCC 6113-3
CBS 243.83
CBS 291.79
SAUCC 6322-5-1*
SAUCC 6151-1
SAUCC 1862-1*
SAUCC 1866-1
CBS 540.92
CPC 25994
CGMCC 3.17929*
GDMCC 3.683
LNU-196
YFCC 8794*
CBS 109067*
CBS 624.94*
BRIP 65649*
CBS 242.90*
CGMCC 3.23525*
CGMCC 3.23524
SAUCC 210782
SAUCC 210781*
SAUCC 1016*
BRIP 75114a
CBS 146.68
CBS 122885*
COAD 3358*
CBS 741.79
SAUCC 211092*
SAUCC 211093
CBS 143499
CBS 143500*
SAUCC 2450-1*
SAUCC 2450-3

ITS

HG934112
KP859044
KP859043
OL897016
OL897017
0L897018

OL897019

OL897020
PP716851

PP716852
PP716853
PP716854
PP716855
KP858994
KP858996
PP695312
PP695313
OR/02567
OR/02568
KP859010
KP859018
KU746690
MT988022
MT988020
OK586161

KP859003

KP858999

MK966337
KP859015
OP103966
OP103965
OM956296
OM956295
MT199884
0Q917075
KP858993

KP859016
0k966954
KP859001

OM956214
OM956215
MH107894
MH107895
OR702569
OR/02570

LSU
MH868531
KP858981
KP858980
OL897058
OL897059

OL897060

OL897061

OL897062
PP716512
PP716513
PP716514
PP716515
PP716516
KP858930
KP858932
PP702043
PP702044
OR702576
OR/02577
KP858946
KP858954
KU746736
MT988024
MT988023
OK586160
KP858939
KP858935
KP858951

OP104016
OP104015
0M959324
0M959323
MT199878
0Q892168
KP858929
KP858952
0k966953

KP858937
OM957532
0M957533
MH107941
MH107942
OR/02578
OR7/02579

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355

GenBank accession number

RPB2
HG934118

ON568988
ON568989
ON568990
PP729053
PP729054
PP729055
PP729056
PP729057
KP859103
KP859105
PP716780
PP716779
OR7/15785
OR/15786
KP859119
KP859127
MW002442
MW002445
OK584019
KP859112
KP859108
KP859124
OP236027
OP236026
OM981154
0M981153
MT510550
0Q889560
KP859102
KP859125
OL310501
KP859110
OM981148
OM981149
MH108003
OR/15787
OR/15788

TUB2
HG934104

ON569069

ON569070

ON569071

ON569072 |

ON569073
PP729058
PP729059

PP729060

PP729061
PP729062
KP859057

KP859059 |

PP716787
PP716788
PP445175
PP445176
KP859073
KP859074
KU746781
MW002441
MW002442
OK556901
KP859066
KP859062
KP859078

OM981147
OM981146
MT435653

KP859056

KP859080
KP859064
OM981141
OM981142

PP445177
PP445178

References

Vu et al. (2019)
Hernandez-Restrepo et al. (2016)

Zhang et al. (2023b)

This study

Hernandez-Restrepo et al. (2016)

This study

Zhang et al. (2023a)

Hernandez-Restrepo et al. (2016)

Zhang et al. (2017)
Lu et al. (2023)

Tang et al. (2022)
Hernandez-Restrepo et al. (2016)

Crous et al. (2020)
Hernandez-Restrepo et al. (2016)
Gao et al. (2022)

Liu et al. (2022)

Huang et al. (2020)
Hernandez-Restrepo et al. (2016)
Crous et al. (2021)
Hernandez-Restrepo et al. (2016)
Liu et al. (2022)

Crous et al. (2018)

Zhang et al. (2023a)

306

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Species

Mi.

ne
Mi
Mi.
Mi
Mi.

Mi

Mi

oqueenslandicum

. nivale
. Nivale var. majus
. Nivale var. nivales

. Novae-zelandiae

. paspali

phyllosaprophyticum

Mi.

Mi

SSS 8

=

SES

S

Selenodriella cubensis

S:

. phragmitis

. poae

i, ratticaudae
ij. rhopalostylidis
i, salmonicolor

i, seminicola

i, shilinense

i, sinense

j. sorghi

|, tainanense

i. trichocladiopsis
Mi

. yunnanense

fertilis

Strain no.

CBS 445.95
CBS 108926*
CBS 116205*

CBS 177.29

CBS 288.50

CPC 29376*

CPC 29693
HK-ML-1371
CBS 138620*

SAUCC 3583-1*
SAUCC 3583-6
CBS 285.71*
CBS 423.78
CGMCC 3.19170*
LGA.

LC 12116

BRIP 68298*
CBS 145125*

NC14-294
CBS 139951*

CPC 26001

DAOM 250161

CGMCC 3.23531*

SAUCC 211097*
SAUCC 211098
SAUCC 3922-1
SAUCC 3922-3
CBS 691.96
CBS 269.76*
CBS 270.76
CBS 623.77*
SAUCC 1011*
SAUCC 1012
CBS 683.96
CBS 772.83

ITS

| KP858997

KP859002
KP859008
MH855031
LT990655
LT990656
KJ569509
KJ569513
OR/02571
OR7/02572
KP859013
KP859012
MH740898
MH740901
MH740902
MW481661
MK442592
MK836110
KP859038
KP859025
KP859034
OP103972
OM956289
OM956290
PP695314
PP695315
KP859000
KP859009
KP858995
KP858998
MT199881
MT199882
KP859053
KP859055

LSU
KP858933
KP858938
KP858944
MH866500
MH868135

OR/02580
OR702581
KP858949
KP858948

MW481666
MK442532
MK836108
KP858974
KP858961
KP858970
OP104022
OM959225
OM959226
PP702045
PP702046
KP858936
KP858945
KP858931
KP858934
MT199875
MT199876
KP858990
KP858992

GenBank accession number

RPB2
KP859106
KP859111
KP859117

LT990641
LT990642

OR715789
OR715790
KP859122
KP859121
MH740906
MH740909
MH740910
MW626890
MK442667
KP859147
KP859134
KP859143
OM981151
OM981152
PP716781
PP716782
KP859109
KP859118
KP859104
KP859107
MT510547
MT510548

TUB2

KP859060 |

KP859065
KP859071

LT990608
LT990609
KJ569514
KJ569518
PP445179
PP445180
KP859077

KP859076 |

MH740914
MH740917
MH740918

KP859101
KP859088

KP859097 |

OP242834

OM981144

OM981145

PP716789

PP716790
KP859063
KP859072

KP859058 |
KP859061
MT435650

MT435651

References

Hernandez-Restrepo et al. (2016)

Vu et al. (2019)

Marin-Felix et al. (2019)

Zhang et al. (2015)

Zhang et al. (2023a)

Hernandez-Restrepo et al. (2016)

Liang et al. (2019)

Crous et al. (2021)

Crous et al. (2019)

Das et al. (2020)
Hernandez-Restrepo et al. (2016)

Gao et al. (2022)

Liu et al. (2022)

This study

Hernandez-Restrepo et al. (2016)

Huang et al. (2020)

Hernandez-Restrepo et al. (2016)
Hernandez-Restrepo et al. (2016)

Notes: Ex-type or ex-epitype strains are marked with “*” and the new species described in this study was marked in bold.

Library construction, quality control and whole-genome sequencing

Library construction and sequencing were carried out by Novogene Co., Ltd.
(Beijing, China). Obtain FASTQ format data, which included sequence infor-
mation and corresponding sequencing quality information (Cock et al. 2010).
Preprocess the raw data that were obtained from the sequencing platform us-
ing fastp (https://github.com/OpenGene/fastp) to obtain clean data for subse-
quent analysis (Chen et al. 2018). Clean data were deposited in the National
Center for Biotechnology Information (NCBI) under BioProject PRJNA1105317.

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355

307

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Genome assembly and annotation

Genome data were assembled using the software SPAdes v 3.12.0 (Bankev-
ich et al. 2012). Genome annotation mainly included three aspects: a. Masking
of repetitive sequences (RepeatMasker version v4.1.4; RepeatModeler v2.0.3,
https://www.repeatmasker.org/); b. Annotation of non-coding RNA (RNAmmer
v1.2; tRNAscan-SE v2.0); c. Annotation of gene structure (RNA-seq prediction:
Trinity v2.14.0, HISAT2 v2.2.1, StringTie v2.2.0; Ab inito prediction: BRAKER2;
Homology protein prediction: GEMoMa v1.9) (Grabherr et al. 2011; Pertea et
al. 2015; Keilwagen et al. 2016, 2018; Bruna et al. 2021). The final genome and
annotation files were integrated using EVM and PASA (Haas et al. 2008, 2011).

Phylogeny

The generated consensus sequences were subjected to Megablast searches to
identify closely related sequences in the NCBI's GenBank nucleotide database
(Zhang et al. 2000). Newly generated sequences in this study were aligned with
related sequences retrieved from GenBank (Table 1) using MAFFT 7 (Katoh et
al. 2019; http://mafft.cbrc.jp/alignment/server/) online service with the default
strategy and corrected manually used MEGA 7. For phylogenetic analyses, we
operated following the methods by Zhang et al. (2023a), single and concatenated
ITS rDNA, LSU, RPB2 and TUB2 sequence alignments were subjected to analysis
by maximum likelihood (ML) and Bayesian Inference (BI) algorithms, respectively.
ML and BI were run on the CIPRES Science Gateway portal (https://www.phylo.
org/, accessed on 30 April 2023) or offline software (ML was operated in Rax-
ML-HPC2 on XSEDE v8.2.12, and BI analysis was operated in MrBayes v3.2.7a
with 64 threads on Linux). For ML analyses, the default parameters were used and
1,000 rapid bootstrap replicates were run with the GTR+G+I model of nucleotide
evolution; BI analysis was performed using a fast bootstrap algorithm with an au-
tomatic stop option (Zhang et al. 2023a). The GTR+I+G model was recommended
for LSU, RPB2, and TUB2, while SYM+I+G was suggested for ITS. The Markov
chain Monte Carlo (MCMC) analysis of the five concatenated genes was con-
ducted over 1,130,000 generations, yielding 22,602 trees. Following the discard
of the initial 5,650 trees generated during the burn-in phase, the remaining trees
were used to compute posterior probabilities in the majority rule consensus trees.

For phylogenomic analyses, the genome sequences were submitted to GenBank
under the accession numbers in Table 2. The final annotated data were processed
to retain the coding protein genes and the longest transcript. Extracted all coding
protein genes to identify gene families and single copy orthologous genes using
OrthoFinder v2.5.5 (https://github.com/davidemms/OrthoFinder), according to the
method by Emms and Kelly (2015, 2019). Multiple sequence alignment was used
ParaAT v1.0 (https://ngdc.cncb.ac.cn/tools/paraat) and merged into supergene us-
ing seqkit v2.7.0 (https://github.com/shenwei356/segkit) (Zhang etal. 2012; Shenet
al. 2016). Phylogenomic analysis was carried out following the methods by Stamat-
akis et al. (2014), using RAXML-NG v1.2.1 (https://github.com/amkozlov/raxml-ng)
with the LG+G8+F model and 100 bootstrap replications. All resulted trees were
plotted using FigTree v. 1.4.4 (http://tree.bio.ed.ac.uk/software/figtree) or ITOL: In-
teractive Tree of Life (https://itol.embl.de/, accessed on 20 October 2023) (Letunic
and Bork 2021) and the layout of the trees was edited in Adobe Illustrator CC 2019.

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 308

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Table 2. BioSample and SRA NCBI number of the taxa used in phylogenomic reconstruction in this study.

Species
Asterophora parasitica
Cryphonectria parasitica
Diaporthe eres
Macroidriella bambusae
Microdochium australe
Mi. bambusae
Mi. bolleyi
Mi. nannuoshanense
Mi. nivale
Mi. phyllosaprophyticum
Mi. trichocladiopsis
Pestalotiopsis fici
Xylaria flabelliformis

Strains BioSample SRA NCBI* References
APO1 SAMN09737569 SRS3956156
EP155 SAMNO2744051 SRS6915724 Crouch et al. 2020
CBS 160.32 SAMN21449118 SRS10459569 Hilario et al. 2022
SAUCC 6792-1 SAMN41099213 SRR28834790 This study
SAUCC 6322-5-1 SAMN41099214 SRR28834789 This study
SAUCC 1862-1 SAMN41099215 SRR28834788 This study
J235TASD1 SAMN04386150 SRS1667728 David et al. 2016
SAUCC 2450-1 SAMN41099216 SRR28834787 This study
F00608 SAMN26062287 SRS14642463 Tsers et al. 2023
SAUCC 3583-1 SAMN41099217 SRR28834786 This study
MPI-CAGE-CH-0230 SAMN06297163 SRS2394902 Mesny et al. 2021
W106-1 SAMN02369365 Wang et al. 2015
G536 SAMN11912834 SRS4852315

Species information described in this study is marked in bold.

Results

Phylogenetic and phylogenomic analyses

A total of 80 isolates representing species within the Microdochiaceae fam-
ily used for phylogenetic analysis. One strain of Cryptostroma corticale (CBS
218 52) was used as an outgroup taxon. The final alignment comprised 3,386
concatenated characters, spanning from positions 1 to 553 (ITS), 554 to 1,827
(LSU), 1,828 to 2,676 (RPB2), and 2,677 to 3,386 (TUB2). The maximum likeli-
hood (ML) optimization likelihood was calculated to be -23041.844775. The
matrix exhibited 1,071 distinct alignment patterns, with 25.57% of characters or
gaps remaining undetermined. MrModelTest suggested that Dirichlet base fre-
quencies be utilized for the ITS, LSU, RPB2, and TUB2 data partitions. The align-
ment exhibited a total of 876 unique site patterns (ITS: 287, LSU: 186, RPB2: 386,
TUB2: 213). The topology of the ML tree corroborated that of the tree obtained
from Bayesian inference; therefore, only the ML tree is depicted (Fig. 1). Based
on the four-gene phylogeny (Fig. 1), the 80 strains were classified into 47 spe-
cies. To enhance the visual appeal and conciseness of the phylogenetic tree,
39 strains were collapsed within it (The complete ML phylogenetic tree is avail-
able in the Suppl. material 5). Among them, five strains (SAUCC 6792-1, SAUCC
6792-2, SAUCC 6792-5, SAUCC 6113-1 and SAUCC 6113-3) identified a new ge-
nus, Macroidriella gen. nov., with solid support (98% MLBV and 1.0 BIPP), and
M. bambusae sp. nov. (SAUCC 6792-1) as the type species. Two strains (SAUCC
6322-5-1 and SAUCC 6151-1) identified as Microdochium australe sp. nov.

We sequenced the genomes of six species in Microdochiaceae for phyloge-
nomic analyses, and downloaded the published genomes of four species from
in NCBI Datasets (https://www.ncbi.nlm.nih.gov/datasets/). Xylaria flabelliformis
G536 was used as an outgroup taxon. Based on 4,909 clusters of orthologous
proteins, the ML tree is depicted (Fig. 2). The phylogenomic tree was divided into
two clades (excepted outgroup), viz, clade 1 (Microdochium nannuoshanense
SAUCC 2450-1, Mi. phyllosaprophyticum SAUCC 3583-1, Mi. australe SAUCC 6322-
5-1 and Mi. bambusae SAUCC 1862-1) and clade 2 (Mi. nivale F00608, Mi. bolleyi
J235TASD1, Mi. trichocladiopsis MPI-CAGE-CH-0230 and Macroidriella bambusae

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<a __ Soap se node with 39 leaves

Microdochium phragmitis CBS 285.71 *

Microdochium phragmitis CBS 423.78

Microdochium rhopalostylidis CBS 145125 *

Microdochium lycopodinum CBS 146.68

Microdochium lycopodinum CBS 125585*

Microdochium chuxiongense YFCC 8794*

Microdochium maculosum COAD 3358*

10071) PAUCC 6151-1
SAUCC 6322-5-1*

-ft00r Microdochium bambusae SAUCC 1866-1

Microdochium bambusae SAUCC 1862-1*

\_ Microdochium indocalami SAUCC 1016*

1001) Microdochium phyllosaprophyticum SAUCC 3583-6

Microdochium phyllosaprophyticum SAUCC 3583-1*

1001) Microdochium nannuoshanense SAUCC 2450-1*

Microdochium nannuoshanense SAUCC 2450-3

SAUCC 3922-3

SAUCC 3922-1 Microdochium sinense

SAUCC 211098

SAUCC 211097*

"| Microdochium salmonicolor NC 14-294

Microdochium miscanthi SAUCC 211092*

Microdochium miscanthi SAUCC 211093

Microdochium fisheri CBS 242.90*

Microdochium hainanense SAUCC 210782

Microdochium hainanense SAUCC 210781*

Microdochium australe sp. nov.

Microdochium
7710.94

91/1

(100/0.9,

(90/0:

go0.93 |/Ariella multiformispora CGMCC 3.20779 *
100/1 let multiformispora GZUIFR 21.924
Idriella multiformispora GZUIFR 21.925
4001) Ariella chlamydospora CGMCC 3.20778 *
Idriella chlamydospora GZUIFR 21.922
hoon (driella lunata CBS 204.56 *
ldriella lunata CBS 177.57

Idriella 4

Figure 1. A maximum likelihood tree was constructed using a combined dataset of ITS, LSU, RPB2, and TUB2 sequence
data. Branch support values, shown as ML/BIPP are indicated above the nodes: MLBV = 70% on the left and BIPP = 0.90
on the right. Ex-type cultures are denoted in bold and marked with an asterisk (*). Strains from the current study are
highlighted in red. The tree was rooted with Cryptostroma corticale (CBS 218.52). The scale bar at the bottom center
represents 0.05 substitutions per site.

= OO ==

Microdochium nannuoshanense SAUCC 2450-1
100) |

Microdochium phyllosaprophyticum SAUCC 3583-1

Microdochium australe SAUCC 6322-5-1

100

Microdochium bambusae SAUCC 1862-1
Microdochium nivale FOO608

Microdochium bolleyi J235TASD1

100

Microdochium trichocladiopsis MP|-CAGE-CH-0230

100

Macroidriella bambusae SAUCC 6792-1

Figure 2. A Maximum Likelihood phylogenomic tree was constructed using a combined 4,909 clusters of orthologous
proteins. Maximum Likelihood bootstrap values (2 70%) are indicated along branches. Genera are highlighted in different
colors. The scale bar at the bottom represents 0.1 substitutions per site.

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Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

SAUCC 6792-1). The branch length of all four strains was < 0.1 in clade 1, indi-
cating that their evolutionary distance was relatively close compared to clade
2 (each strain’s branch was > 0.1). Due to limited genomic data, Macroidriella
bambusae (SAUCC 6792-1) was not individually clustered, but the evolutionary
distance of Macroidriella bambusae is relatively far compared to other species.

Annotations and comparative analysis

After structural annotation of the genomic data, we conducted a statistical
summary, including, number of genes, total number of cds, total number of ex-
ons, total number of introns, total cds length, total exon length and total intron
length (Suppl. material 2). Due to the limited genomic data available for Micro-
dochiaceae, we will conduct gene family analysis by comparing the self-tested
data of the new genus (Macroidriella) with genomic data from the orders of
Diaporthales (Cryphonectria parasitica EP155 and Diaporthe eres CBS 160.32),
Xylariales (Pestalotiopsis fici W106-1 and Xylaria flabelliformis G536), and the
Basidiomycota (Asterophora parasitica APO1). The intersections of gene fam-
ily among the six representative strains (s 6) are 3431, the maximum number
(508) of gene family intersections between Macroidriella bambusae and Micro-
dochium trichocladiopsis, and the minimum number (4) of gene family inter-
sections between Macroidriella bambusae and Asterophora parasitica (Fig. 3a).
The intersections of gene family among the seven representative strains are
3,291, the unique number of genes in Asterophora parasitica was 513 (maxi-
mum), the unique number of genes in Macroidriella bambusae was 42 (mini-
mum) (Fig. 3b). We have presented the number of single-copy genes, multi-co-
py genes and so on for the seven representative strains (Fig. 3c).

Taxonomy

Macroidriella Z.X. Zhang, J. W. Xia & X.G. Zhang, gen. nov.
MycoBank No: 853699

Type species. Macroidriella bambusae Z. X. Zhang, J. W. Xia & X. G. Zhang.

Etymology. Referring to the composed of “Macro-” and “-idriella” (Similar in
morphology to /driella and bigger than /driella in conidia).

Description. Genus of Microdochiaceae. Endogenic on diseased leaves of
Bambusaceae sp. Sporodochia yellowish brown, slimy. Conidiophores are in-
distinct and often reduced to conidiogenous cells. Conidiogenous cells are
straight or slightly branched, smooth, curved, mono- or polyblastic, terminal, hy-
aline, septate, cylindrical and ampulliform. Conidia are solitary, hyaline, lunate,
curved, mooned, multi-guttulate, apex rounded, base usually flattened. Sexual
morphs were not observed, chlamydospores were not observed.

Notes. In the phylogenetic tree (Fig. 1), Macroidriella is allied to /driella,
Microdochium and Selenodriella, but forms a separate lineage with good sta-
tistical support (98% MLBV and 1.0 BIPP). In morphology, the conidia of Mac-
roidriella are predominantly lunate and curved, unlike the elliptical conidia of
Microdocium, suggesting a genus of its own, because it is similar to /driella
in morphology (but the conidia of Macroidriella are longer than /driella), both
being lunate conidia, it is named Macroidriella gen. nov.

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 311

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Asterophora parasitica
Cryphonectria parasitica
Diaporthe eres

Macroidriella bambusae
Microdochium trichocladiopsis
Pestalotiopsis fici

Xylaria flabelliformis

a 3431

aie

2774

L]
[|
[|
fa]
L

gene family intersection size

7500. 5000 2500 is}
gene family set number

b C 15000

a Other orthologs
|| Unique paralogs
a Multi-copy orthologs
ay Single-copy orthologs

Gene number

Figure 3. Gene family analysis of Macroidriella a UpSet plot of six strains, showing the intersection counts between
different strains in the form of a bar graph b petal plot of seven strains, the center of the petal represents the number of
shared genes c bar chart of homologous genes for each strain.

Macroidriella bambusae Z.X. Zhang & X.G. Zhang, sp. nov.
MycoBank No: 853712
Fig. 4

Type. CHINA, Hainan Province, Danzhou City: Hainan tropical botanical garden,
on diseased leaves of Bambusaceae sp., 15 October 2023, Z. X. Zhang (HMAS
352974, holotype), ex-holotype living culture SAUCC 6792-1.
Etymology. Referring to the species name of the host plant Bambusaceae sp.
Description. Endogenic on diseased leaves of Bambusaceae sp. Mycelia are
superficial and immersed, 2—3.5 um wide, branched, membranous and hyaline.

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Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Figure 4. Macroidriella bambusae (HMAS 352974, holotype) aa leaf of Bambusaceae sp. b, c colonies on PDA from above
and below after 14 days d colony overview e, f conidiogenous cells and conidia g, h conidia. Scale bars: 10 um (eh).

Sporodochia yellowish brown, slimy. Conidiophores are indistinct and often re-
duced to conidiogenous cells. Conidiogenous cells are straight or slightly curved,
10.4-15 x 1.7-—2.8 um, mono- or polyblastic, terminal, hyaline, septate, cylindri-
cal and smooth. Conidia are solitary, hyaline, lunate, curved, mooned, 16.5-
21.7 x 2-2.8 um, multi-guttulate, apex rounded, base usually flattened. Sexual
morphs were not observed, chlamydospores were not observed, see Fig. 4.

Culture characteristics. Cultures incubated on PDA at 25 °C in darkness,
reaching 63-70 mm diam., had a growth rate of 4.5—5.0 mm/day after 14 days,
with moderate aerial mycelia, the center and edges are milky white, with a yel-
low-brown color in the middle, and sporodochia are observed.

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Additional material studied. CHINA, Hainan Province, Danzhou City, Hainan trop-
ical botanical garden, on diseased leaves of Bambusaceae sp., 15 October 2023, Z.
X. Zhang (HSAUP 6792-2), living culture SAUCC 6792-2; ibid, (HSAUP 6792-5), living
culture SAUCC 6792-5; on dead leaves, 15 October 2023, Z. X. Zhang (HSAUP 6113-
1), living culture SAUCC 6113-1; ibid., (HSAUP 6113-3), living culture SAUCC 6113-3.

Notes. Phylogenetic analyses showed that Macroidriella bambusae formed
an independent clade (Fig. 1), and closely related to /driella multiformispora (lu-
nate, curved-shaped conidia) and Microdochium bolleyi. The Ma. bambusae was
distinguished from |. multiformispora (CGMCC 3.20779) by 60/520, 22/1222,
74/848 and 57/710 base-pair differences, from Mi. bolleyi (CBS 540.92) by
40/514, 19/765, 138/850 and 51/710 base pairs in ITS, LSU, RPB2 and TUB2 se-
quences, respectively. Morphologically, Ma. bambusae (16.5-21.7 x 2—2.8 pm)
longer than /. multiformispora (8.5-13.5 x 1.0-2 um) and Mi. bolleyi (5-8.7 x
1.6-2.3 um) in conidia. Therefore, we describe this fungus as a novel species.

Microdochium australe Z.X. Zhang, & X.G. Zhang, sp. nov.
MycoBank No: 853695
Fig..5

Type. CHINA, Hainan Province, Jianfengling National Forest Park, on diseased
leaves of Phragmites australis, 13 October 2023, Z. X. Zhang (HMAS 352973,
holotype), ex-holotype culture SAUCC 6322-5-1.

Etymology. Referring to the species name of the host plant Phragmites australis.

Description. Endogenic on diseased leaves of Phragmites australis. Mycelia are
superficial and immersed, 3-3.3 um wide, branched, membranous and hyaline.
Sporodochia black, aggregative or solitary. Conidiophores are indistinct and often
reduced to conidiogenous cells. Conidiogenous cells are straight or slightly curved,
15.4-23.5x2.8-—4 um, terminal, hyaline, septate, ampulliform or obpyriform, smooth.
Conidia are solitary, hyaline, straight to slight curved, oblong to ellipsoid, 11.3-16.1
x 2.5-3.7 um, multi-guttulate, (2)3-septate, apex rounded, base usually flattened.
Sexual morphs were not observed, chlamydospores were not observed, see Fig. 5.

Culture characteristics. Cultures incubated on PDA at 25 °C in darkness,
reaching 73-76 mm diam., had a growth rate of 5.2—5.4 mm/day after 14 days,
with moderate aerial mycelia, milky white to grey-white, with regular margin,
and sporodochia are observed, reverses black to brown in the centre, with grey-
white and regular margin.

Additional material studied. CHINA, Hainan Province, Jianfengling National
Forest Park, on saprophytic leaves, 13 October 2023, Z. X. Zhang (HSAUP 6151-
1), living culture SAUCC 6151-1.

Notes. Phylogenetic analyses showed that Microdochium australe sp. nov.
formed an independent clade closely related to Microdochium bambusae and Mi-
crodochium indocalami (Fig. 1). Mi. australe was distinguished from Mi. bambusae
(SAUCC 1862-1) by 47/503, 2/836, 56/848 and 17/710 base pair differences, from
Mi. bambusae and Mi. indocalami (SAUCC 1016) by 52/503, 2/848, 44/840 and
17/708 base pairs in ITS, LSU, RPB2 and TUB2 sequences, respectively. Morpho-
logically, Mi. australe (11.3-16.1 x 2.5-3.7 um, (2)3-septate) differs from Mi. bam-
busae (13.0-17 x 2.5-3.5 um, aseptate) and Mi. indocalami in conidia (13-15.5 x
3.5-5.5 um, 3-septate), and, therefore, we described this fungus as a novel species.

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Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Figure 5. Microdochium australe (HMAS 352973, holotype) a a leaf of Phragmites australis b, c colonies on PDA from above
and below after 14 days d colony overview e, f conidiogenous cells and conidia g, h conidia. Scale bars: 10 um (eh).

Microdochium sinense S.B. Liu, X.Y. Liu, Z. Meng & X.G. Zhang, J. Fungi
2022, 8, 577.
Fig. 6

Material examined. CHINA, Hainan Province, Jianfengling National Forest Park,
on diseased leaves of Phragmites australis, 12 April 2023, Z. X. Zhang (HSAUP
3922-1), living culture SAUCC 3922-1; ibid., (HSAUP 3922-3), living culture
SAUCC 3922-3.

Description. Endogenic on diseased leaves of Phragmites australis.
Mycelia are superficial and immersed, 2.1-2.9 um wide, branched, membra-
nous and hyaline. Conidia are solitary, hyaline, straight, oblong to ellipsoid,

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Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

e a a fT
Figure 6. Microdochium sinense a diseased symptoms on a leaf of Phragmites australis b, c colonies on PDA from above
and below after 14 days d conidiomata on PDA e, f conidia. Scale bars: 10 um (e-f).

12.3-15 x 3.5-5.6 um, multi-guttulate, apex rounded, base usually flattened.
Conidiophores were not observed, chlamydospores were not observed, sexual
morphs were not observed, see Fig. 6.

Culture characteristics. Cultures incubated on PDA at 25 °C in darkness,
reach-ing 72-76 mm diam., had a growth rate of 5.1-5.4 mm/day after 14
days, with moder-ate aerial mycelia, milky white to grey-white, with irregular
margin, reverses light brown in the centre, with grey-white and regular margin.

Notes. Phylogenetic analyses of four combined genes (ITS, LSU, RPB2 and
TUB2) showed that SAUCC 3922-1 and SAUCC 3922-3 clustered with the type
collection of Microdochium sinense with strong support (Fig. 1). We, therefore,
identified the isolated strains (SAUCC 3922-1 and SAUCC 3922-3) as Mi. sin-
ense. Morphologically, the conidia of the both (newly isolated and type) were
similar (12.3-15 x 3.5-5.6 vs. 11.5-19.34 x 2.8-5.4 um).

Discussion

The establishment of the family Microdochiaceae by Hernandez-Restrepo et
al. (2016) to encompass the clade consisting of /driella, Microdochium, and
Selenodriella within the Xylariales highlights the importance of phylogenetic
analysis in understanding the evolutionary relationships among fungi. This new
classification helps to better organize and categorize fungal species based on
their genetic relatedness and morphological characteristics (Hernandez-Re-
strepo et al. 2016; Liang et al. 2019; Huang et al. 2020; Liu et al. 2022; Lu et al.

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Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

2023; Zhang et al. 2023a). In the recent study, nine strains isolated from two
host plants, Phragmites australis and Bambusaceae sp., were introduced into
a new genus, Macroidriella and two new species, Macroidriella bambusae and
Microdochium australe. The Global Biodiversity Information Facility (GBIF) cur-
rently hosts 1,594 georeferenced records of Microdochiaceae species world-
wide (https://www.gbif.org/, accessed on April 30, 2024). The distribution of
this family is predominantly in the United States, Europe, and Oceania, with
fewer occurrences in Asia.

In the recent study of the family, Microdochium emerged as a prominent re-
search focus, with 12 species of this genus documented across five Provinces
(Guizhou, Hainan, Henan, Shandong, and Yunnan) since the beginning of the
21st century in China (Zhang et al. 2015; Liang et al. 2019; Huang et al. 2020;
Gao et al. 2022; Liu et al. 2022; Tang et al. 2022). Microdochium species have
been identified on a variety of host families (10 families), with over half of the
fungi associated with Poaceae plants. In contrast, /driella and Selenodriella
have been less extensively studied, with /driella having only two reported spe-
cies since the turn of the 21% century. Through the joint analysis of multiple
gene fragments and genomes, the position of new taxa can be better deter-
mined, especially through phylogenomic analyses, which was provided with
more robust support values. Comparative analysis will help us determine the
position of the Macroidriella genus on the evolutionary tree and its relationship
with other fungi. By comparing the genomic data of different fungi, we can iden-
tify common gene families and infer their evolutionary relationships. Through
comparative genomic analysis, it can be observed that Macroidriella has 42
unique single-copy orthologous genes. Asterophora shares only 4 single-copy
orthologous genes with Macroidriella, which also indicates that their relation-
ship is very distant (belonging to different fungal phyla).

This study represents a pioneering effort in Microdochiaceae as it integrates
multi-gene fragments with genomic data to unveil the phylogenetic relation-
ships within the family. By combining these diverse datasets, a comprehen-
sive understanding of the evolutionary history of Microdochiaceae is achieved,
shedding new light on its genetic landscape and evolutionary dynamics.

Acknowledgements
We would like to express our gratitude to Jie Zhang, a master’s student at Shan-

dong Agricultural University, for her assistance in this study.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

This research was supported by National Natural Science Foundation of China (nos.
32100016, 32270024, U2002203 and 32370001).

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 317

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Author contributions

Sampling, molecular biology analysis: Zhao-Xue Zhang and Yu-Xin Shang; fungal isola-
tion: Yu-Xin Shang and Jin-Jia Zhang; description and phylogenetic analysis: Meng-Yu-
an Zhang; microscopy: Yun Geng; writing—original draft preparation: Zhao-Xue Zhang;
writing—review and editing, Ji-Wen Xia and Xiu-Guo Zhang. All authors read and
approved the final manuscript.

Author ORCIDs

Zhao-Xue Zhang ® https://orcid.org/0000-0002-4824-9716
Ji-Wen Xia © https://orcid.org/0000-0002-7436-7249

Data availability

All of the data that support the findings of this study are available in the main text or
Supplementary Information.

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Supplementary material 1
The PCR primers, sequence and cycles used in this study

Authors: Zhao-Xue Zhang, Yu-Xin Shang, Meng-Yuan Zhang, Yun Geng, Ji-Wen Xia,
Xiu-Guo Zhang

Data type: docx

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/mycokeys.106.127355.suppl1

Supplementary material 2
GenBank accession number of the taxa used in phylogenetic reconstruction

Authors: Zhao-Xue Zhang, Yu-Xin Shang, Meng-Yuan Zhang, Yun Geng, Ji-Wen Xia,
Xiu-Guo Zhang

Data type: docx

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/mycokeys.106.127355.suppl2

Supplementary material 3
The sequence of phylogenetic analysis

Authors: Zhao-Xue Zhang, Yu-Xin Shang, Meng-Yuan Zhang, Yun Geng, Ji-Wen Xia,
Xiu-Guo Zhang

Data type: txt

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/mycokeys.106.127355.suppl3

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 394

Zhao-Xue Zhang et al.: Phylogenomics, taxonomy and morphological characters of the Microdochiaceae

Supplementary material 4
The sequence of phylogenomic analysis

Authors: Zhao-Xue Zhang, Yu-Xin Shang, Meng-Yuan Zhang, Yun Geng, Ji-Wen Xia,
Xiu-Guo Zhang

Data type: txt

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/mycokeys.106.127355.suppl4

Supplementary material 5

The complete ML phylogenetic tree

Authors: Zhao-Xue Zhang, Yu-Xin Shang, Meng-Yuan Zhang, Yun Geng, Ji-Wen Xia,
Xiu-Guo Zhang

Data type: pdf

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/mycokeys.106.127355.suppl5

MycoKeys 106: 303-325 (2024), DOI: 10.3897/mycokeys.106.127355 395